The AngioPhase™ Kit incorporates a patented technology in which human endothelial cells are co-cultured with other human cells in a specially designed medium. The endothelial cells initially form small islands within the culture matrix. They subsequently begin to proliferate and then enter a migratory phase during which they move through the matrix to form threadlike tubule structures. These gradually join up (by 1 - 2 weeks) to form a network of anastomosing tubules which closely resembles the capillary bed found in the CAM assay. The tubules stain positive for von Willebrand’s Factor, PECAM 1 and ICAM-2. They are also shown to secrete a matrix containing type IV collagen.

The AngioPhase™ Kit assay is responsive to known micro and macro molecular inhibitors and stimulators of angiogenesis and so, unlike some other models, measures both positive and negative effects on angiogenesis. It yields reproducible dose response curves permitting comparison of different treatment regimes and product concentrations.

In addition to primary cells, seeding and growth media (with associated supplements), the AngioPhase™ Kit also includes:

• Validated antibodies and reagents for tubule visualization.
• Validated positive control (VEGF).
• Validated negative control (Suramin).

The AngioPhase™ Kit contains all of the cells and reagents necessary to successfully complete a 24 well angiogenesis assay. The kit contains a cryogenically preserved ampule of matched cells, 24 well tissue culture plate, medium, supplements, control compounds and all staining reagents.

The AngioPhase™ Kit is designed so that test compounds, conditioned media or tissue explants can be added to the culture at any time from the onset of vasculogenesis continuing through to advanced angiogenesis. The resulting effect on tubule formation can then be measured either manually or with the aid of AngioSys Image Analysis Software.

Control wells that receive no treatment other than medium changes form extensive networks of branching tubules over a period of one to two weeks. This allows both angiogenic inhibitors and angiogenic stimulators to be assessed. The necessary medium changes are included in the kit.

Positive and negative control compounds are included in the AngioPhase™ Kit which consist of validated concentrated stock solutions of VEGF (2μg/ml, positive control) and Suramin (1mM, negative control) ready to add to medium.